Abstract
The concerns regarding microplastics and nanoplastics pollution stimulate studies on the uptake and biodistribution of these emerging pollutants in vitro. Atomic force microscopy in nanomechanical PeakForce Tapping mode was used here to visualise the uptake and distribution of polystyrene spherical microplastics in human skin fibroblast. Particles down to 500 nm were imaged in whole fixed cells, the nanomechanical characterization allowed for differentiation between internalized and surface attached plastics. This study opens new avenues in microplastics toxicity research.
Highlights
Microplastics pollution is currently regarded to be among severe environmental threats [1], it is clear that most, if not all, habitats are polluted with microplastic to a certain extent [2]
It is clear that polystyrene microbeads are not fully representative as other types of microplastics [5], especially the environmental specimens, which may and do have a range of sizes, shapes and chemical composition, in the first approximation the polystyrene microbeads have been employed in a number of the microplastics-related studies, especially aimed at elucidation of biological activity in vitro
Developing effective methods to visualise and identify micro- and nanoplastics is of paramount importance in modern bioanalytical chemistry [12], since understanding of pollution extent in natural habitats and toxicity pathways in cells and organisms is impossible without quantitative identification of polymeric solids [13], which are exceptionally diverse and hard to isolate from complex biological matrices
Summary
Microplastics pollution is currently regarded to be among severe environmental threats [1], it is clear that most, if not all, habitats are polluted with microplastic to a certain extent [2]. Developing effective methods to visualise and identify micro- and nanoplastics is of paramount importance in modern bioanalytical chemistry [12], since understanding of pollution extent in natural habitats and toxicity pathways in cells and organisms is impossible without quantitative identification of polymeric solids [13], which are exceptionally diverse and hard to isolate from complex biological matrices Spectroscopy techniques, such as FTIR or Raman microspectroscopy [14] are routinely used to identify microscale plastics in environmental samples, which are rarely used to investigate mechanisms of microplastics toxicity due to their complex chemistry, shape and size. We hypothesized that using AFM in nanomechanical mode (that is where the image acquisition is followed by quantitative measurements of such parameters as modulus, adhesion, etc.) might be applicable for detection of microplastics taken up by human cells if the latter are chemically fixed and dried, following the routine AFM sample preparation protocol In this case, the solid spherical microplastics will be an easy target for the tip of AFM probe, having spatial geometry and mechanical properties different from the adjacent cellular compartments. In this Communication we demonstrate the feasibility of our technique, using human skin fibroblasts primary cells, submicron polystyrene microbeads and PeakForce Tapping nanomechanical AFM
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